卤醇脱卤酶是一种多功能生物催化剂，通过分子内亲核取代反应高效、高选择地催化邻卤醇合成环氧化物及其逆反应，用来合成光学纯的环氧化物及β-取代醇等一类高价值中间体，极具应用潜能。目前，对上述酶促反应机理及其结构基础并未进行深入研究，不利于该酶的高效改造研究。鉴于该酶具有独特的内源荧光特性和其底物具有特征红外吸收，本项目在前期研究基础上，拟采用稳态-瞬态荧光光谱、停流荧光、FTIR、同时辅以飞秒二维红外技术，从多时间尺度、多结构层面阐述卤醇脱卤结构动态性的酶学功能，重点阐述酶调控酶催化活性的构象变化的结构基础及底物多样性的调控机制；结合蛋白质改造技术和理论计算等手段对相关研究进行验证、丰富和加以完善，进而建立卤醇脱卤酶催化活性及其底物多样性的精细调控网络。相关研究不仅有利于丰富相关酶促反应的理论基础，还将极大地促进该酶高效改造及其应用研究。

Halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) displays a great potential in the synthesis of chiral building blocks-epoxides and a series of β-substituted alcohols for pharmaceuticals. However, the kinetic mechanism of the enzyme, especially its related structural dynamics has not been investigated in detail, which largely limits its industrial application. Taking the advantage of the fact that HheC exhibits a novel instinct fluorescence property and its substrates (nucleophiles) owning a unique character of IR spectrum, HheC represents a suitable model for exploring the relationship between enzyme function and the related structural dynamics. Thus, in this project, techniques such as steady-state and transient fluorescence spectroscopy, stopped-flow technique, FTIR and femtosecond 2D IR will be used to explore the structural dynamics of a functional important conformational change occurring in the dehalogenation reaction of HheC and the nucleophile promiscuity mechanism of the enzyme. In addition, enzyme mutagenesis techniques, MD simulation and QM/MM will be used as supplement tools for further proving. In the end, a relationship between the structural dynamics profiles and the enzyme activity will be built through data analysis and mapping. The results will greatly facilitate not only the theoretical understanding of the enzyme kinetics, but also the enzyme application in the field of chiral drug synthesis.